Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseases
In this study, we isolated and cultured canine and feline 3D corneal organoids. Samples derived from corneal limbal epithelium from one canine and one feline patient were obtained by enucleation after euthanasia. Stem cell isolation and organoid culture were performed by culturing organoids in Matri...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2022.1050467/full |
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| author | Leila Bedos Hannah Wickham Vojtech Gabriel Christopher Zdyrski Rachel A. Allbaugh Dipak Kumar Sahoo Lionel Sebbag Lionel Sebbag Jonathan P. Mochel Jonathan P. Mochel Karin Allenspach Karin Allenspach Karin Allenspach |
| author_facet | Leila Bedos Hannah Wickham Vojtech Gabriel Christopher Zdyrski Rachel A. Allbaugh Dipak Kumar Sahoo Lionel Sebbag Lionel Sebbag Jonathan P. Mochel Jonathan P. Mochel Karin Allenspach Karin Allenspach Karin Allenspach |
| author_sort | Leila Bedos |
| collection | DOAJ |
| description | In this study, we isolated and cultured canine and feline 3D corneal organoids. Samples derived from corneal limbal epithelium from one canine and one feline patient were obtained by enucleation after euthanasia. Stem cell isolation and organoid culture were performed by culturing organoids in Matrigel. Organoids were subsequently embedded in paraffin for further characterization. The expression of key corneal epithelial and stromal cell markers in canine and feline organoids was evaluated at the mRNA level by RNA-ISH and at the protein level by immunofluorescence (IF) and immunohistochemistry (IHC), while histochemical analysis was performed on both tissues and organoids using periodic-acid Schiff (PAS), Sirius Red, Gomori's Trichrome, and Colloidal Iron stains. IF showed consistent expression of AQP1 within canine and feline organoids and tissues. P63 was present in canine tissues, canine organoids, and feline tissues, but not in feline organoids. Results from IHC staining further confirmed the primarily epithelial origin of the organoids. Canine and feline 3D corneal organoids can successfully be cultured and maintained and express epithelial and stem cell progenitor markers typical of the cornea. This novel in vitro model can be used in veterinary ophthalmology disease modeling, corneal drug testing, and regenerative medicine. |
| first_indexed | 2024-04-12T14:22:00Z |
| format | Article |
| id | doaj.art-8ae8c25abfea4fa2b79e05a78a603745 |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-04-12T14:22:00Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Veterinary Science |
| spelling | doaj.art-8ae8c25abfea4fa2b79e05a78a6037452022-12-22T03:29:33ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692022-11-01910.3389/fvets.2022.10504671050467Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseasesLeila Bedos0Hannah Wickham1Vojtech Gabriel2Christopher Zdyrski3Rachel A. Allbaugh4Dipak Kumar Sahoo5Lionel Sebbag6Lionel Sebbag7Jonathan P. Mochel8Jonathan P. Mochel9Karin Allenspach10Karin Allenspach11Karin Allenspach12Department of Veterinary Clinical Sciences, Iowa State University, Ames, IA, United StatesSMART Lab, Department of Biomedical Sciences, Iowa State University, Ames, IA, United StatesSMART Lab, Department of Biomedical Sciences, Iowa State University, Ames, IA, United StatesSMART Lab, Department of Biomedical Sciences, Iowa State University, Ames, IA, United StatesDepartment of Veterinary Clinical Sciences, Iowa State University, Ames, IA, United StatesDepartment of Veterinary Clinical Sciences, Iowa State University, Ames, IA, United StatesDepartment of Veterinary Clinical Sciences, Iowa State University, Ames, IA, United StatesKoret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, IsraelSMART Lab, Department of Biomedical Sciences, Iowa State University, Ames, IA, United States3D Health Solutions Inc., Ames, IA, United StatesDepartment of Veterinary Clinical Sciences, Iowa State University, Ames, IA, United StatesSMART Lab, Department of Biomedical Sciences, Iowa State University, Ames, IA, United States3D Health Solutions Inc., Ames, IA, United StatesIn this study, we isolated and cultured canine and feline 3D corneal organoids. Samples derived from corneal limbal epithelium from one canine and one feline patient were obtained by enucleation after euthanasia. Stem cell isolation and organoid culture were performed by culturing organoids in Matrigel. Organoids were subsequently embedded in paraffin for further characterization. The expression of key corneal epithelial and stromal cell markers in canine and feline organoids was evaluated at the mRNA level by RNA-ISH and at the protein level by immunofluorescence (IF) and immunohistochemistry (IHC), while histochemical analysis was performed on both tissues and organoids using periodic-acid Schiff (PAS), Sirius Red, Gomori's Trichrome, and Colloidal Iron stains. IF showed consistent expression of AQP1 within canine and feline organoids and tissues. P63 was present in canine tissues, canine organoids, and feline tissues, but not in feline organoids. Results from IHC staining further confirmed the primarily epithelial origin of the organoids. Canine and feline 3D corneal organoids can successfully be cultured and maintained and express epithelial and stem cell progenitor markers typical of the cornea. This novel in vitro model can be used in veterinary ophthalmology disease modeling, corneal drug testing, and regenerative medicine.https://www.frontiersin.org/articles/10.3389/fvets.2022.1050467/fullorganoidcorneacaninefelinestem cellcomparative ophthalmology |
| spellingShingle | Leila Bedos Hannah Wickham Vojtech Gabriel Christopher Zdyrski Rachel A. Allbaugh Dipak Kumar Sahoo Lionel Sebbag Lionel Sebbag Jonathan P. Mochel Jonathan P. Mochel Karin Allenspach Karin Allenspach Karin Allenspach Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseases Frontiers in Veterinary Science organoid cornea canine feline stem cell comparative ophthalmology |
| title | Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseases |
| title_full | Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseases |
| title_fullStr | Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseases |
| title_full_unstemmed | Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseases |
| title_short | Culture and characterization of canine and feline corneal epithelial organoids: A new tool for the study and treatment of corneal diseases |
| title_sort | culture and characterization of canine and feline corneal epithelial organoids a new tool for the study and treatment of corneal diseases |
| topic | organoid cornea canine feline stem cell comparative ophthalmology |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2022.1050467/full |
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